以卵巢幹細胞培養出成熟卵細胞顛覆更年期後無法製造卵子之生理限制

最新研究顯示未來可能分離卵巢組織內之卵子幹細胞

植入去免疫力之老鼠卵細胞中以體外培養出成熟之卵細胞

未來此研究進一步成熟發展可能顛覆傳統女性於更年期後就無法製造卵子之生理限制

http://blogs.nature.com/spoonful/2012/02/video-stem-cell-discovery-puts-women%e2%80%99s-reproduction-on-fertile-grounds.html

Nat Med. 2012 Feb 26;18(3):413-21. doi: 10.1038/nm.2669.

Oocyte formation by mitotically active germ cells purified from ovaries of reproductive-age women.

White YA, Woods DC, Takai Y, Ishihara O, Seki H, Tilly JL.

Source

Vincent Center for Reproductive Biology, Massachusetts General Hospital Vincent Department of Obstetrics and Gynecology, Massachusetts General Hospital, Boston, Massachusetts, USA.

Abstract

Germline stem cells that produce oocytes in vitro and fertilization-competent eggs in vivo have been identified in and isolated from adult mouse ovaries. Here we describe and validate a fluorescence-activated cell sorting-based protocol that can be used with adult mouse ovaries and human ovarian cortical tissue to purify rare mitotically active cells that have a gene expression profile that is consistent with primitive germ cells. Once established in vitro, these cells can be expanded for months and can spontaneously generate 35- to 50-μm oocytes, as determined by morphology, gene expression and haploid (1n) status. Injection of the human germline cells, engineered to stably express GFP, into human ovarian cortical biopsies leads to formation of follicles containing GFP-positive oocytes 1-2 weeks after xenotransplantation into immunodeficient female mice. Thus, ovaries of reproductive-age women, similar to adult mice, possess rare mitotically active germ cells that can be propagated in vitro as well as generate oocytes in vitro and in vivo.

(a) Immunofluorescence analysis of Ddx4 expression (green; with a blue DAPI counterstain) in young adult mouse ovaries using antibodies against the N (NH₂) or the C (COOH) terminus of the protein (scale bars, 50 μm). (b) Immunomagnetic sorting of dis…

(a,b) Representative histological appearance of adult ovarian tissue used for the isolation of human (a) and mouse (b) OSCs. Scale bars, 100 μm. (c,d) Morphology of viable cells isolated by FACS based on cell-surface expression of DDX4 (human, left)…

(a,b) Examples of growing follicles containing GFP-negative (a) and GFP-positive (b) (brown; with a blue hematoxylin counterstain) oocytes in ovaries of wild-type mice injected with GFP-expressing OSCs 5–6 months before analysis. (c) Examples of ovul…

(a–d) Assessment of OSC proliferation by dual detection of Ddx4 or DDX4 expression (green) and BrdU incorporation (red) in mouse (a,b) and human (c,d) OSCs maintained in MEF-free cultures. Scale bars, 30 μm. (e) Typical growth curve for MEF-free cult…